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Regenerating and enhancing development of muscle tissue

a technology of muscle tissue and regenerative mechanism, which is applied in the direction of biocide, drug composition, peptide/protein ingredients, etc., can solve the problems of increasing the impaired muscle regenerative mechanism, but without completely salvaging the altered phenotype, etc., and achieves the effects of regenerating muscle tissue, and enhancing muscle tissue developmen

Inactive Publication Date: 2009-11-19
SBARRO HEALTH RES ORG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0009]The present invention pertains to a method for regenerating muscle tissue and enhancing development of muscle tissue, comprising administering a vector encoding cdk9-55 to a patient in need thereof. The vector may be an adenoviral vector or a retroviral vector. The vector may be administered intra-arterially, intravenously, or intramuscularly. The muscle tissue may be smooth muscle, cardiac muscle, and / or skeletal muscle. The present invention also pertains to a method for regenerating muscle tissue or enhancing development of muscle tissue, comprising co-administering a vector encoding cdk9-55 and Cyt2a, MyoD, or at least one muscle regulatory factor to the patient in need of muscle tissue regeneration. The present invention also pertains to a method for regenerating muscle tissue or enhancing development of muscle tissue, comprising administering to a patient a composition comprising cdk9-55 proteins. Further, at least one muscle regulatory factor and / or adjuvant may be administered with cdk9-55.

Problems solved by technology

Many different therapeutic approaches have been developed, giving rise to an increase in impaired muscle regenerative mechanisms but without completely rescuing the altered phenotype.

Method used

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  • Regenerating and enhancing development of muscle tissue
  • Regenerating and enhancing development of muscle tissue
  • Regenerating and enhancing development of muscle tissue

Examples

Experimental program
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Effect test

example 1

Cell Culture

[0067]C2C12 cells were grown in DMEM supplemented with 20% FBS (GM) or with 2% HS (DM).

[0068]Single muscle fibers with associated satellite cells were isolated as described in Rosenblatt et al., In Vitro Cell. Dev. Biol. Anim. 31, 773-779, 1995. Briefly, the hind limb muscles were digested for 60 minutes at 37° C. in 0.2% collagenase I (Sigma-Aldrich) and triturated with a wide-bore pipette. Single myofibers were then cultured in 6-well plates (10 fibers / well) pre-coated with ECM gel (Sigma-Aldrich). Fiber cultures were grown in DMEM supplemented with 10% HS and 0.5% chick embryo extract (MP Biomedicals, Solon, Ohio). Three days later, the fibers were removed, and proliferation of detached cells was induced by DMEM supplemented with 20% FBS, 10% HS, and 1% chick embryo extract. After 4-5 days, the cells were allowed to differentiate by DMEM with 2% HS and 0.5% chick embryo extract.

example 2

Muscle Regeneration and Immufluorescence Studies

[0069]Quadriceps and tibialis muscles from C57BL6J mice and 22-month Desmin / nls-LacZ mice were injected with 30 μl of 10 μM cardiotoxin. Mice were anesthetized before cervical dislocation and muscle tissue was separated from bone and most connective tissue and immediately frozen in liquid nitrogen. Frozen sections were fixed in 4% paraformaldehyde 10 min on ice, and then stained with X-gal or pre-incubated in PBS containing 1% BSA, 1:100 goat serum for 1 h at RT, and processed as described in Musarò et al., Nature Genetics 27, 195-200, 2001. Antibodies specific for MyoD, cdk9, Desmin and MHC were used. Nuclei were visualized using Hoechst staining.

example 3

Plasmids and In Vivo Muscle Transfection by Electric Field

[0070]The construct pcDNA3-cdk9DN-HATag expressing dominant negative cdk9 has been described (Simone et al., Oncogene 21, 4137-4148, 2002). In vivo experiments were carried out as described in Dona et al., Biochemical and Biophysical Research Communications 312, 1132-1138, 2003. Briefly, injured quadriceps and tibialis muscles from C57BL6J mice and 22-month Desmin / nls-LacZ mice were exposed by a short incision and DNA (0.06-25 mg) in 50 ml of 0.9% NaCl was injected with a Hamilton syringe in a proximal to distal direction. Then, a pair of spatula-like electrodes (0.5 cm wide, 2 cm long) were placed at each side of the muscle and electric pulses were delivered. Five electric pulses with a fixed pulse duration of 20 ms and an interval of 200 ms were delivered using an electric pulse generator (Electro Square porator ECM 830, BTX, San Diego, Calif.). The ratio of applied voltage to electrode distance was 50 V / cm.

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Abstract

Muscle tissue regeneration is one of the most important homeostatic processes of adult skeletal muscle, which after development retains the capacity to regenerate in response to different type of stimuli, including a direct trauma or neurological dysfunction; atrophy; and genetic defects. The present invention pertains to cdk9-55 and its ability to regenerate muscle tissue and enhance development. Cdk9-55 is specifically induced upon satellite cell differentiation and is necessary for the gene expression reprogramming required to complete the regeneration process.

Description

TECHNICAL FIELD OF THE INVENTION[0001]The present invention relates to a method for regenerating muscle tissue and enhancing muscle tissue development comprising administering cdk9-55.BACKGROUND OF THE INVENTION[0002]Tissue regeneration is one of the most important homeostatic processes of adult skeletal muscle, which after development retains the capacity to regenerate in response to different type of stimuli, including a direct trauma or neurological dysfunction and genetic defects (Huard et al., J. Bone Joint Surg. Am. 84-A, 822-32, 2002). The regenerative process is sustained by adult myogenic precursors, a population of quiescent mononucleated reserve cells, termed satellite cells (Charge et al., Physiol. Rev. 84, 209-238, 2004). Upon exposure to signals from the damaged environment, satellite cells are activated and start proliferating. At the molecular level, this process is characterized by the rapid up-regulation of two muscle regulatory factors (MRFs), Myf5 and MyoD. After...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/45A61K31/711A61P21/00A61K48/00
CPCA61K31/711A61K38/45C12N2799/027A61K48/005C12N2799/022A61K48/0016A61P21/00
Inventor GIORDANO, ANTONIOGIACINTI, CRISTINA
Owner SBARRO HEALTH RES ORG
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